CN109880747A - A kind of preparation method of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps - Google Patents
A kind of preparation method of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps Download PDFInfo
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Abstract
The present invention relates to the preparation methods of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps.Metabolism regulators are added by the different cultivation stages in aweto (Hirsutella sinensis), intervene the metabolism of Hirsutella sinensis, the metabolic pathway for especially intervening routine culture object and wild cordyceps Difference of Metabolism object, makes the metabolin for the hirsutella sinensis fungal manually cultivated closer to wild cordyceps.Adjusted result is shown, Hirsutella sinensis mycelia metabolin and wild cordyceps metabolin after Metabolism regulation cluster together, its finger-print is almost the same simultaneously, shows that wild cordyceps can be replaced by the artificial culture of Hirsutella sinensis of Metabolism regulation of the present invention.Hirsutella sinensis mycelia product of the invention be by indoor fermenting and producing, can batch production whole year production, do not influenced by adverse circumstances such as the low temperature in plateau of wild cordyceps, do not destroy ecological environment, had a vast market.
Description
Technical field
The invention belongs to the culture preparation technical fields of biological products, and in particular to a kind of culture of Hirsutella sinensis mycelia
Method.
Background technique
Cordyceps sinensis is a kind of famous Chinese traditional herbs in China, has the applicating history in over thousands of year, neat with ginseng, pilose antler
Name.It is aweto and a kind of entomogenous fungi complex that parasitic insect is formed, the polypide part including being full of mycelia (sclerotium)
With the fructification part grown by head.Modern pharmacological studies have shown that cordyceps sinensis can adjust immune system, anti-aging,
Blood pressure lowering adjusts endocrine, eliminates fatigue, anti-curing oncoma, acute or chronic hepatitis, ephritis, a variety of pharmacology such as antibacterial, antiviral
Activity is worth with important medical and health care.Since natural cordyceps growing environment is special, and vulnerable to external environment
It influences, in addition artificial predation formula acquisition, so that natural cordyceps quantity is constantly reduced, natural cordyceps resource has been at present
In the state for being on the verge of exhaustion.In order to solve the contradiction that resource exhaustion and demand but constantly rise, people strive to find the worm summer in winter
The substitute of grass.
Existing Jinshuibao and Bailing capsule etc. more than 40 is in relation to substitution obtained by Chinese caterpillar fungus bacterial filament or fermentation material at present
Product are invested in market, although however, these substitutes all claim that its ingredient and cordyceps sinensis are close, the strain largely used
It is not real aweto, strain used is related to more than more than ten categories 20, such as Paecilomyces hepiali chen
(Paecilomyces), cephalo it is mould (Cephalosporiumsp.), Paecilomyces sinensis (Paecilomyces sinensis), bat
Bat moth column it is mould (Scytalidium hepiali), Soil taxonomy (Tolypocladium sinense), the golden spore of China it is mould
(Chrysosporium sinense), Mortierella hepiali (Mortierella hepiali), cylinder bacterium
(Scytalidiumsp.) etc..With the development of modern molecular biology and the application of the other a variety of confirmation means of combination, mesh
Preceding Hirsutella sinensis (Hirsutella sinensis) it has been confirmed to be real aweto, and recognized by the whole world.
We have discovered that the resulting mycelia of non-aweto or culture, ingredient and wild cordyceps difference
It is very big, even if using the artificial culture mycelia of real aweto (Hirsutella sinensis) or culture, ingredient and wild winter
Worm summer grass also has different.In fact, the not only artificial culture mycelia of Hirsutella sinensis or culture, ingredient and wild winter
Worm summer grass has a different, the different parts and different stages of growth of wild cordyceps, and ingredient is also not quite identical.
Summary of the invention
To keep the artificial culture of Hirsutella sinensis (mycelium) consistent with the metabolin of wild cordyceps, Chinese quilt is realized
The hair artificial culture of spore (mycelium) can replace wild cordyceps, and the present invention is to aweto (the Chinese quilt manually cultivated
Hair spore) Metabolism regulation has been carried out, make the ingredient and wild cordyceps ingredient one of the artificial culture of Hirsutella sinensis (mycelium)
It causes.The present invention is directed to provide the preparation method of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps.
Steps are as follows for the preparation manipulation of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps:
(1) level-one solid spawn is prepared
By the Hirsutella sinensis isolated and purified (Hirsutella sinensis) slant strains are inoculated in Solid media for plates
In, every slant strains connect 2 9cm plates；Then it in 17~20 DEG C of temperature, cultivates 20~30 days, obtains level-one solid spawn；
The solid medium is potato culture；
(2) secondary liquid strain is prepared
Level-one solid spawn is inoculated in fluid nutrient medium, each 9cm plate strain connects 200mL fluid nutrient medium；After inoculation
In 17~20 DEG C of temperature, 160~200 revs/min of revolving speed, shaken cultivation 10~20 days, secondary liquid strain is obtained；
(3) three grade fermemtation object is prepared
Secondary liquid strain is accessed in fermentor by 5~10% volume ratios, the fermentation medium liquid amount 50~70% of fermentor,
Ventilatory capacity 1:1(v/v by volume), pressure 0.1MP, under the conditions of 17~20 DEG C of temperature, it is added 100 after culture 5~15 days~
No. bis- metabolism regulators of 500mg/L No.1 metabolism regulators and 50~250mg/L；10~50mg/L is added after culture 10~20 days
No. three metabolism regulators；Continue culture 20~40 days, obtains three grade fermemtation object；
The No.1 regulator is zinc sulfate or zinc chloride；
No. two regulators are sub- oleoyl glycerophosphorylethanolamines or sub- oleoyl serine phosphorylation ester；
No. three regulators are plant sheath amine alcohol or 1- phosphoric acid sheath amine alcohol；
(4) the artificial culture of Hirsutella sinensis is prepared
By the centrifuge separation of three grade fermemtation object, mycelia is filtered out, 100 DEG C of temperature or less dry, pulverize to get Hirsutella sinensis people
Work culture, the artificial culture of Hirsutella sinensis are pale powder；The ingredient of the artificial culture of Hirsutella sinensis and natural winter
The worm summer composition of grass is consistent, and the two has similar finger-print, and wherein mannitol content is greater than 3.5%, adenosine content and is greater than
0.04%, sterol content is greater than 0.3%.
The preparation manipulation technical solution further limited is as follows:
In step (1), the solid medium: by 200.0 g liquor of potato, 20.0 g of glucose, 10.0 g of maltose, egg
White 10.0 g of peptone, 10.0 g of yeast powder, magnesium sulfate (MgSO4) 1.5 g, potassium dihydrogen phosphate (KH2PO4) 3.0 g, 20.0 g of agar,
1.0 L are added water to, adjust pH value to be made for 6.5 with the sodium hydroxide (NaOH) of concentration 1.0M.
In step (2), the fluid nutrient medium: by 20.0 g of glucose, 30.0 g of dried silkworm chrysalis meal, 20.0 g of yeast powder, sulphur
Sour magnesium (MgSO4) 0.5g, potassium dihydrogen phosphate (KH2PO4) 1.0 g are sufficiently mixed, add water to mend to 1000mL, and with 1.0 M's of concentration
Sodium hydroxide (NaOH) adjusts pH value to be made for 6.5.
In step (3), the fermentation medium is by 15.0~20.0 g of glucose, 20.0~30.0 g of dried silkworm chrysalis meal, yeast
15.0~20.0 g of powder, magnesium sulfate (MgSO4) 0.3~0.5g, potassium dihydrogen phosphate (KH2PO4) 0.5~1.0 g is sufficiently mixed, adds
Water is mended to 1000mL, and adjusts pH value to be made for 6.5 with the sodium hydroxide of 1.0 M of concentration (NaOH).
In step (4), the centrifuge separation condition: 5000~10000 revs/min of revolving speed, 5~15 minutes time.
It is described to be filtered into 50~150 meshes in step (4).
Advantageous effects of the invention are embodied in following several aspects:
1, the high-resolution LC-MS fingerprint of the artificial culture of Hirsutella sinensis of the invention (mycelium) and wild cordyceps
Map is completely the same, shows that their ingredients are consistent, can really replace wild cordyceps.Pass through metabolism group method, Wo Menfa
The key component and its metabolic pathway for having showed artificial cultured mycelia and wild cordyceps Difference of Metabolism, pass through to have found
The method of Metabolism regulation makes manually to cultivate mycelia and wild fructification metabolin is more consistent.Referring to Fig. 1 and Fig. 2, clustering
Show that manually cultivating mycelia (quadrangle point in Fig. 1) and wild cordyceps (triangle point in Fig. 1) before Metabolism regulation does not have
It gets together, shows that its metabolin is inconsistent, the artificial culture mycelia (triangle point in Fig. 2) after Metabolism regulation and wild winter
Worm summer grass (the quadrangle point in Fig. 2) is got together, the artificial culture mycelia metabolin after showing Metabolism regulation and wild winter worm
Summer grass is consistent.A kind of commercially available aweto culture product (five-pointed star in Fig. 1) as control cannot be poly- with cordyceps sinensis
To together.Obviously, the Hirsutella sinensis mycelium after Metabolism regulation is more suitable for the substitute of wild cordyceps.
Metabolin it was found that: the mannitol of Hirsutella sinensis mycelium and wild cordyceps after Metabolism regulation,
Nucleosides, peptides and sterol constituents are all consistent, and the main component of the two finger-print is consistent (see Fig. 3,4).
2, the artificial culture of Hirsutella sinensis of the invention (mycelium), can the life of batch production whole year by indoor fermenting and producing
It produces, is not influenced by adverse circumstances such as the low temperature in plateau of wild cordyceps.Wild cordyceps are needed at 4000 meters of height above sea level or more
High mountain growth, not only bad environments, acquire Harvest time that is difficult, and only having a wheat harvesting period every year.It is artificial to cultivate Chinese coat
Spore is cultivated for 17 ~ 20 DEG C indoors, does not need extreme environment, also, by vaccinization and fermentation, it can be achieved that produce, often throughout the year
It has product, and production efficiency is greatly improved.
3, the artificial culture of Hirsutella sinensis of the invention (mycelium) production cost is low, every kilogram of cost 100 yuan with
Under, the hundreds of thousands of members of wild cordyceps per kilogram are far below, also far below the per kilogram number of the indoor bionic cultivation of cordyceps sinensis
Ten thousand yuan of costs.
4, the artificial culture of Hirsutella sinensis (mycelium) is produced using artificial fermentation and replaces wild cordyceps, can protected
Wild cordyceps place of production ecological environment avoids resource exhaustion and deterioration of grasslands caused by excessively excavating.
Detailed description of the invention
Fig. 1 be Metabolism regulation before the artificial culture of Hirsutella sinensis (mycelium) and wild cordyceps and one kind it is commercially available
The metabolism dendrogram of product.
Fig. 2 be Metabolism regulation after the artificial culture of Hirsutella sinensis (mycelium) and wild cordyceps and one kind it is commercially available
The metabolism dendrogram of product.
Fig. 3 is the Hirsutella sinensis mycelium metabolin fingerprint image after wild cordyceps and Metabolism regulation.
Fig. 4 is the Hirsutella sinensis mycelium metabolin fingerprint image after Metabolism regulation.
Fig. 5 is the artificial culture metabolin fingerprint image of Hirsutella sinensis in embodiment 1 after Metabolism regulation.
Fig. 6 is the artificial culture metabolin fingerprint image of Hirsutella sinensis in embodiment 2 after Metabolism regulation.
Fig. 7 is the artificial culture metabolin fingerprint image of Hirsutella sinensis in embodiment 3 after Metabolism regulation.
Specific embodiment
With reference to the accompanying drawing, the present invention is further described by embodiment.
Embodiment 1:
Steps are as follows for the preparation manipulation of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps:
1) strain selects
From Qinghai acquire cordyceps sinensis (Ophiocordyceps sinensis) Hirsutella sinensis isolated on stroma
(Hirsutella sinensis).
) strain culturing
Cultural method is that liquid is mixed admittedly, and specific process is as follows:
2.1) level-one solid spawn is prepared
The slant strains isolated and purified are inoculated in the solid medium of 9 cm culture dishes, solid medium is by potato
200.0 g liquors, 20.0 g of glucose, 10.0 g of maltose, 10.0 g of peptone, 10.0 g of yeast powder, magnesium sulfate (MgSO4)
1.5 g, potassium dihydrogen phosphate (KH2PO4) 3.0 g, 20.0 g of agar, 1.0 L are added water to, with the sodium hydroxide of concentration 1.0M
(NaOH) pH value is adjusted to be made for 6.5.17 DEG C of incubators are put into, incubation time is 20 days, obtains level-one solid spawn；
2.2) secondary liquid shaking flask strain is prepared
In 500mL triangular flask be packed into 200ml fluid nutrient medium, fluid nutrient medium by 20.0 g of glucose, 30.0 g of dried silkworm chrysalis meal,
20.0 g of yeast powder, magnesium sulfate (MgSO4) 0.5g, potassium dihydrogen phosphate (KH2PO4) 1.0 g are sufficiently mixed, add water to mend to 1000mL,
And pH value is adjusted to be made for 6.5 with the sodium hydroxide of 1.0 M of concentration (NaOH).With 1.2 Kg/cm2High pressure sterilization 30 minutes, to cold
When but to room temperature, the solid spawn of 1 culture dish is inoculated into 200mL fluid nutrient medium, constant-temperature shaking incubator is placed in,
It is cultivated 10 days under conditions of 17 DEG C of temperature, 160 revs/min, obtains secondary liquid strain；
2.3) three grade fermemtation object is prepared
By secondary liquid seed by 5% volume ratio access 1000mL triangular flask, the liquid amount of fermentation medium is 400mL, juxtaposition
Under the conditions of constant-temperature shaking incubator, 17 DEG C of temperature, 160 revs/min, 100 mg/L No.1 Metabolism regulations are added after 5 days in culture
No. bis- metabolism regulators of agent and 50 mg/L；Continue to cultivate 10 days addition No. tri- metabolism regulators of 10 mg/L, continues constant temperature oscillation
Culture 20 days, obtains three grade fermemtation object；
Fermentation medium is by 15.0 g of glucose, 20.0 g of dried silkworm chrysalis meal, 15.0 g of yeast powder, magnesium sulfate (MgSO4) 0.3g, phosphorus
Acid dihydride potassium (KH2PO4) 0.5 g is sufficiently mixed, add water to mend to 1000mL, and adjust pH with the sodium hydroxide of 1.0 M of concentration (NaOH)
Value is made for 6.5；No.1 regulator is zinc sulfate, and No. two regulators are sub- oleoyl glycerophosphorylethanolamines, and No. three regulators are
Plant sheath amine alcohol.
) prepare the artificial culture of Hirsutella sinensis
Three grade fermemtation object is centrifuged 5 minutes under the conditions of 5000 revs/min of revolving speed, the bacterium for obtaining centrifugation under the conditions of -40 DEG C
Silk is freeze-dried up to the artificial culture of Hirsutella sinensis, and the artificial culture of Hirsutella sinensis is pale powder；Chinese coat
The high-resolution LC-MS finger-print of the artificial culture of spore and the high-resolution LC-MS finger-print one of wild cordyceps
It causes, the peak as it can be seen that on the LC-MS map of the artificial culture of Hirsutella sinensis and wild cordyceps is compared by Fig. 5 and Fig. 3
Position (retention time) and peak intensity are substantially the same, and show the ingredient type and content of the artificial culture of Hirsutella sinensis all and naturally
The composition of cordyceps sinensis is consistent.Only content of fatty acid is relatively low for the artificial culture of Hirsutella sinensis, but fatty acid is not cordyceps sinensis
Effective component.
Embodiment 2:
Steps are as follows for the preparation manipulation of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps:
1) strain selects
From Qinghai acquire cordyceps sinensis (Ophiocordyceps sinensis) Hirsutella sinensis isolated on stroma
(Hirsutella sinensis)；
2) strain culturing
Cultural method is that liquid is mixed admittedly, and specific process is as follows:
2.1) level-one solid spawn is prepared
The slant strains isolated and purified are inoculated in the solid medium of 9 cm culture dishes (culture medium are as follows: 200.0 g of potato
Liquor, 20.0 g of glucose, 10.0 g of maltose, 10.0 g of peptone, yeast powder 10.0 g, MgSO4 1.5 g、KH2PO4
3.0 g, 20.0 g of agar, 1.0 L are added water to, is to 20 DEG C of incubators, incubation time 6.5), is put into the NaOH tune pH of 1.0M
30 days, obtain level-one solid spawn；
2.2) secondary liquid shake-flask seed is prepared
It is packed into 200ml fluid nutrient medium in 500mL triangular flask, 20.0 g of glucose, dried silkworm chrysalis meal are contained in 1000mL culture medium
30.0 g, 20.0 g of yeast powder, MgSO4 0.5g, KH2PO41.0 g, after each component mixing plus water is mended to 1000mL, and with 1.0
The NaOH tune pH value of M is 6.5, obtains fluid nutrient medium；With 1.2 Kg/cm2High pressure sterilization 30 minutes, when being cooled to room temperature,
The level-one solid spawn of 1 culture dish is inoculated into 200mL fluid nutrient medium, constant-temperature shaking incubator, temperature 20 are placed in
DEG C, it is cultivated 20 days under conditions of 180 revs/min, obtains secondary liquid strain；
2.3) three grade fermemtation object is prepared
By secondary liquid strain by 10% volume ratio access fermentor, the fermentation medium liquid amount of fermentor is tank volume
50%；Contain 20.0 g of glucose, 30.0 g of dried silkworm chrysalis meal, yeast powder 20.0 g, MgSO in every liter of fermentation medium4 0.5g、KH2PO4
1.0 g, pH value 6.5.Ventilatory capacity 1:1(v/v by volume), pressure 0.1MP, 20 DEG C of temperature are added 500mg/ after culture 15 days
No. bis- metabolism regulators of L No.1 metabolism regulators and 250mg/L；Continue to cultivate 20 days addition No. tri- metabolism regulators of 50mg/L.
No.1 regulator is zinc chloride.No. two regulators are sub- oleoyl serine phosphorylation ester.No. three regulators are 1- phosphoric acid sheath amine alcohol.
) prepare the artificial culture of Hirsutella sinensis
After three grade fermemtation object constant-temperature shaking culture 40 days of fermentor, 10000 revs/min are centrifuged 15 minutes, in 100 DEG C of conditions
The lower mycelia forced air drying for obtaining centrifugation is up to the Hirsutella sinensis mycelium after Metabolism regulation.The high-resolution liquid matter of the mycelia
Be combined finger-print it is consistent with wild cordyceps, compared by Fig. 6 and Fig. 3 as it can be seen that the artificial culture of Hirsutella sinensis with it is wild
Peak position (retention time) and peak intensity on the LC-MS map of cordyceps sinensis are substantially the same, and show that Hirsutella sinensis is manually trained
The ingredient type and content for supporting object are all consistent with the composition of natural cordyceps.Only content of fatty acid is relatively low, but fatty acid is not
The effective component of cordyceps sinensis.
Embodiment 3:
Steps are as follows for the preparation manipulation of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps:
1) strain selects
From Qinghai acquire cordyceps sinensis (Ophiocordyceps sinensis) Hirsutella sinensis isolated on stroma
(Hirsutella sinensis)；
2) strain culturing
Cultural method is that liquid is mixed admittedly, and specific process is as follows:
2.1) level-one solid spawn is prepared
The slant strains isolated and purified are inoculated in the solid medium of 9 cm culture dishes, solid medium are as follows: potato
200.0 g liquors, 20.0 g of glucose, 10.0 g of maltose, 10.0 g of peptone, yeast powder 10.0 g, MgSO4 1.5 g、
KH2PO43.0 g, 20.0 g of agar, 1.0 L are added water to, with the NaOH tune pH to 6.5 of 1.0M；18.5 DEG C of incubators are put into, are trained
Supporting the time is 25 days, obtains level-one solid spawn；
2.2) secondary liquid shake-flask seed is prepared
200ml fluid nutrient medium is packed into 500mL triangular flask；Contain 20.0 g of glucose, dried silkworm chrysalis meal in every 1000mL culture medium
30.0 g, 20.0 g of yeast powder, MgSO4 0.5g, KH2PO41.0 g, after each component mixing plus water is mended to 1000mL, and with 1.0
The NaOH tune pH value of M is 6.5；With 1.2 Kg/cm2High pressure sterilization 30 minutes, when being cooled to room temperature, by the one of 1 culture dish
Grade solid spawn is inoculated into 200mL fluid nutrient medium, is placed in constant-temperature shaking incubator, and 18.5 DEG C, 180 revs/min of temperature
Under conditions of cultivate 15 days, obtain secondary liquid strain；
2.3) three grade fermemtation object is prepared
By secondary liquid strain by 8% volume ratio access fermentor；The fermentation medium liquid amount of fermentor is tank volume
60%；Contain 17.5 g of glucose, 25.0 g of dried silkworm chrysalis meal, yeast powder 17.5 g, MgSO in every liter of fermentation medium4 0.4g、KH2PO4
0.75 g, pH value 6.5.Ventilatory capacity 1:1(v/v by volume), pressure 0.1MP, cultivate 7 days under the conditions of 18.5 DEG C of temperature and be added
No. bis- metabolism regulators of 300 mg/L No.1 metabolism regulators and 150 mg/L；Continue addition in 15 days 30mg/L tri- metabolism of culture
Regulator.No.1 regulator is zinc chloride.No. two regulators are sub- oleoyl serine phosphorylation ester.No. three regulators are 1- phosphoric acid sheath
Amine alcohol.
) prepare the artificial culture of Hirsutella sinensis
After three grade fermemtation object constant-temperature shaking culture 30 days of fermentor, crosses 75 meshes and filter out mycelia, by bacterium under the conditions of -40 DEG C
Silk is freeze-dried up to the Hirsutella sinensis mycelium after Metabolism regulation.The high-resolution LC-MS finger-print of the mycelia and open country
Raw cordyceps sinensis is consistent, is compared by Fig. 7 and Fig. 3 as it can be seen that the liquid matter of the artificial culture of Hirsutella sinensis and wild cordyceps joins
With on map peak position (retention time) and peak intensity be substantially the same, show the artificial culture of Hirsutella sinensis ingredient type and
Content is all consistent with the composition of natural cordyceps.
Claims (6)
1. the preparation method of a kind of ingredient and the artificial culture of the consistent Hirsutella sinensis of natural cordyceps, it is characterised in that behaviour
Steps are as follows for work:
(1) level-one solid spawn is prepared
By the Hirsutella sinensis isolated and purified (Hirsutella sinensis) slant strains are inoculated in Solid media for plates
In, every slant strains connect 2 9cm plates；Then it in 17~20 DEG C of temperature, cultivates 20~30 days, obtains level-one solid spawn；
The solid medium is potato culture；
(2) secondary liquid strain is prepared
Level-one solid spawn is inoculated in fluid nutrient medium, each 9cm plate strain connects 200mL fluid nutrient medium；After inoculation
In 17~20 DEG C of temperature, 180 revs/min of revolving speed, shaken cultivation 10~20 days, secondary liquid strain is obtained；
(3) three grade fermemtation object is prepared
Secondary liquid strain is accessed in fermentor by 5~10% volume ratios, the fermentation medium liquid amount 50~70% of fermentor,
Ventilatory capacity 1:1(v/v by volume), pressure 0.1MP, under the conditions of 17~20 DEG C of temperature, it is added 100 after culture 5~15 days~
No. bis- metabolism regulators of 500mg/L No.1 metabolism regulators and 50~250mg/L；10~50mg/L is added after culture 10~20 days
No. three metabolism regulators；Continue culture 20~40 days, obtains three grade fermemtation object；
The No.1 regulator is zinc sulfate or zinc chloride；
No. two regulators are sub- oleoyl glycerophosphorylethanolamines or sub- oleoyl serine phosphorylation ester；
No. three regulators are plant sheath amine alcohol or 1- phosphoric acid sheath amine alcohol；
(4) the artificial culture preparation of Hirsutella sinensis
By the centrifuge separation of three grade fermemtation object, mycelia is filtered out, 100 DEG C of temperature or less dry, pulverize to get Hirsutella sinensis people
Work culture, the artificial culture of Hirsutella sinensis are pale powder, the ingredient of the artificial culture of Hirsutella sinensis and natural winter
The worm summer composition of grass is consistent, and wherein mannitol content is greater than 3.5%, adenosine content and is greater than 0.04%, sterol content greater than 0.3%.
2. preparation method according to claim 1, it is characterised in that: in step (1), the solid medium: by Ma Ling
200.0 g liquor of potato, 20.0 g of glucose, 10.0 g of maltose, 10.0 g of peptone, 10.0 g of yeast powder, magnesium sulfate
(MgSO4) 1.5 g, potassium dihydrogen phosphate (KH2PO4) 3.0 g, 20.0 g of agar, 1.0 L are added water to, with the hydroxide of concentration 1.0M
Sodium (NaOH) adjusts pH value to be made for 6.5.
3. preparation method according to claim 1, it is characterised in that: in step (2), the fluid nutrient medium: by grape
20.0 g of sugar, 30.0 g of dried silkworm chrysalis meal, 20.0 g of yeast powder, magnesium sulfate (MgSO4) 0.5g, potassium dihydrogen phosphate (KH2PO4) 1.0 g fill
Divide mixing, water is added to mend to 1000mL, and adjusts pH value to be made for 6.5 with the sodium hydroxide of 1.0 M of concentration (NaOH).
4. preparation method according to claim 1, it is characterised in that: in step (3), the fermentation medium is by glucose
15.0~20.0 g, 20.0~30.0 g of dried silkworm chrysalis meal, 15.0~20.0 g of yeast powder, magnesium sulfate (MgSO4) 0.3~0.5g, phosphorus
Acid dihydride potassium (KH2PO4) 0.5~1.0 g is sufficiently mixed, add water to mend to 1000mL, and with the sodium hydroxide of 1.0 M of concentration
(NaOH) pH value is adjusted to be made for 6.5.
5. preparation method according to claim 1, it is characterised in that: in step (4), the centrifuge separation condition: revolving speed
5000~10000 revs/min, the time 5~15 minutes.
6. preparation method according to claim 1, it is characterised in that: described to be filtered into 50~150 mesh in step (4)
Sieve.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1796539A (en) * | 2004-12-24 | 2006-07-05 | 青海月王青藏药业有限责任公司 | Ferment for producing aweto in large scale and technique for processing power of fungus |
| CN101000331A (en) * | 2007-01-05 | 2007-07-18 | 李绍平 | Method for investigating quality of characterization natural cordyceps sinensis character |
| CN103609329A (en) * | 2013-11-05 | 2014-03-05 | 昆山市康乐虫草专业合作社 | Cordyceps militaris culturing method capable of improving cordycepin content |
| CN105886412A (en) * | 2016-05-12 | 2016-08-24 | 中山大学 | Liquid fermentation culture medium for cordyceps sinensis |
-
2019
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1796539A (en) * | 2004-12-24 | 2006-07-05 | 青海月王青藏药业有限责任公司 | Ferment for producing aweto in large scale and technique for processing power of fungus |
| CN101000331A (en) * | 2007-01-05 | 2007-07-18 | 李绍平 | Method for investigating quality of characterization natural cordyceps sinensis character |
| CN103609329A (en) * | 2013-11-05 | 2014-03-05 | 昆山市康乐虫草专业合作社 | Cordyceps militaris culturing method capable of improving cordycepin content |
| CN105886412A (en) * | 2016-05-12 | 2016-08-24 | 中山大学 | Liquid fermentation culture medium for cordyceps sinensis |
Non-Patent Citations (3)
| Title |
|---|
| XUANWEI ZHOU ET AL.: ""Cordyceps Fungi: Natural Products, Pharmacological Functions and Developmental Products"", 《J PHARM PHARMACOL.》 * |
| 李淑林等: "培养时间相关的中国被毛孢代谢产物初步分析", 《菌物学报》 * |
| 毛先兵等: "采用氨基酸补料分批培养策略促进蛹虫草生产虫草素的研究", 《重庆中草药研究》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113061534A (en) * | 2021-02-22 | 2021-07-02 | 广东省食用菌行业协会 | Method for preserving hirsutella sinensis strain |
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